Zhang R, Pan X, Huang Z, Weber GF, Zhang GOsteopontin enhances the expression and activity of MMP-2 via the SDF-1/CXCR4 axis in hepatocellulat carcinoma cell lines. PLoS One 6(8): e23831

Institut de Génomique Fonctionnelle de Lyon, France
PLoS ONE (Impact Factor: 3.23). 08/2011; 6(8):e23831. DOI: 10.1371/journal.pone.0023831
Source: PubMed


Osteopontin, SDF-1α, and MMP-2 are important secreted molecules involved in the pathophysiology of human hepatocellular carcinoma (HCC). This study investigates the effect of the SDF-1α/CXCR4 axis on expression and activity of MMP-2 induced by osteopontin.
The expression of CXCR4, SDF-1α, MMP-2 and their associated cellular signaling cascades, involving Akt and MAP Kinases, were determined by Western blotting. The activities of MMP-2 and MMP-9 were assayed by gel zymography. The role of the osteopontin receptors integrin α(v)β₃ and CD44v6 was evaluated using neutralizing antibodies. We also established CXCR4-deficient SMMC7721 cell lines by transfection with miRNA-CXCR4 plasmids and determined cell invasion activity in a transwell assay.
In comparison with untreated cells, recombinant human osteopontin (rhOPN) up-regulated CXCR4, SDF-1α, and MMP-2 expression about 5-, 4-, and 6-fold on the protein levels through binding to integrin α(v)β₃ and CD44v6 in hepatocellular carcinoma cells (SMMC7721 and HepG2). Inhibition of the SDF-1α/CXCR4 axis down-regulated the rhOPN-induced MMP-2 expression and activity. rhOPN also activated Akt, p38 and JNK. Down-regulation of CXCR4 decreased the rhOPN-induced invasion in SMMC7721 cells.
These results indicate that rhOPN up-regulates MMP-2 through the SDF-1α/CXCR4 axis, mediated by binding to integrin α(v)β₃ and CD44v6 and activating the PI-3K/Akt and JNK pathways in HepG2 and SMMC7721 cells. Therefore, the osteopontin-SDF-1α/CXCR4-MMP-2 system may be a new therapeutic target for treating HCC progression.

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    • "MMPs are regulated precisely during synthesis and secretion , at the time of activation of the pro - enzyme and / or by localisation , clearance and inhibition of the active enzyme ( Ra and Parks , 2007 ) . With respect to activation , previous studies have shown that MMP - 2 is activated by osteopontin ( Zhang et al , 2011 ) , reactive oxygen species ( ROS ; Svineng et al , 2008 ) , TGF - b2 ( Baumann et al , 2009 ) and TIMP - 2 ( Itoh et al , 2001 ; Rosenthal and Matrisian , 2006 ) , among other mechanisms . It is particularly interesting , therefore , that osteopontin is a key constituent of the SASP ( Coppé et al , 2008 ) and is known to promote epithelial tumour development ( Pazolli et al , 2009 ) . "
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    ABSTRACT: Background: Previous studies have demonstrated that senescent cancer-associated fibroblasts (CAFs) derived from genetically unstable oral squamous cell carcinomas (GU-OSCC), unlike non-senescent CAFs from genetically stable carcinomas (GS-OSCC), promoted keratinocyte invasion in vitro in a paracrine manner. The mechanism by which this occurs is unclear. Methods: Previous work to characterise the senescent-associated secretory phenotype (SASP) has used antibody arrays, technology that is limited by the availability of suitable antibodies. To extend this work in an unbiased manner, we used 2D gel electrophoresis and mass spectroscopy for protein identification. Matrix metalloproteinases (MMPs) were investigated by gelatin zymography and western blotting. Neutralising antibodies were used to block key molecules in the functional assays of keratinocyte adhesion and invasion. Results: Among a variety of proteins that were differentially expressed between CAFs from GU-OSCC and GS-OSCC, MMP-2 was a major constituent of senescent CAF-CM derived from GU-OSCC. The presence of active MMP-2 was confirmed by gelatine zymography. MMP-2 derived from senescent CAF-CM induced keratinocyte dis-cohesion and epithelial invasion into collagen gels in a TGF-β-dependent manner. Conclusions: Senescent CAFs from GU-OSCC promote a more aggressive oral cancer phenotype by production of active MMP-2, disruption of epithelial adhesion and induction of keratinocyte invasion.
    British Journal of Cancer 08/2014; 111(6). DOI:10.1038/bjc.2014.438 · 4.84 Impact Factor
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    • "OPN also activates Nrf2 signaling, resulting in enhanced heme oxygenase expression and cell migration in glioma cells (Lu et al., 2012). In HCC cells OPN promotes TGF-β1 mediated hepatic stellate cell activation (Xiao et al., 2012) and also upregulates CXCR4, SDF-1α, and MMP-2 expression through binding to integrin αvβ3 and CD44v6 (R. Zhang et al., 2011). Functionally, OPN is required for vascular mimicry in HCC cells (Liu et al., 2011). "
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    ABSTRACT: Osteopontin (OPN) is a multifunctional cytokine that impacts cell proliferation, survival, drug resistance, invasion, and stem like behavior. Due to its critical involvement in regulating cellular functions, its aberrant expression and/or splicing is functionally responsible for undesirable alterations in disease pathologies, specifically cancer. It is implicated in promoting invasive and metastatic progression of many carcinomas. Due to its autocrine and paracrine activities OPN has been shown to be a crucial mediator of cellular cross talk and an influential factor in the tumor microenvironment. OPN has been implicated as a prognostic and diagnostic marker for several cancer types. It has also been explored as a possible target for treatment. In this article we hope to provide a broad perspective on the importance of OPN in the pathophysiology of cancer.
    Matrix biology: journal of the International Society for Matrix Biology 03/2014; 37. DOI:10.1016/j.matbio.2014.03.001 · 5.07 Impact Factor
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    • "To validate that Huh7.5 cells induce above signaling pathway through the interaction of OPN at the cell surface receptors, mock (Huh7.5) cells were incubated with recombinant human OPN (rhOPN from R&D Systems) (50 nM) as described earlier [39]. The immunoblot results showed the increased phosphorylation/activation of FAK, Akt, N-cadherin and Src but decreased expression of E-cadherin in presence of rhOPN by western blot analysis (Fig. 6C, lane 2, 4). "
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    ABSTRACT: Osteopontin (OPN) is a secreted phosphoprotein which has been linked to tumor progression and metastasis in a variety of cancers including hepatocellular carcinoma (HCC). Previous studies have shown that OPN is upregulated during liver injury and inflammation. However, the role of OPN in hepatitis C virus (HCV)-induced liver disease pathogenesis is not known. In this study, we determined the induction of OPN, and then investigated the effect of secreted forms of OPN in epithelial to mesenchymal transition (EMT), migration and invasion of hepatocytes. We show the induction of OPN mRNA and protein expression by HCV-infection. Our results also demonstrate the processing of precursor OPN (75 kDa) into 55 kDa, 42 kDa and 36 kDa forms of OPN in HCV-infected cells. Furthermore, we show the binding of secreted OPN to integrin αVβ3 and CD44 at the cell surface, leading to the activation of downstream cellular kinases such as focal adhesion kinase (FAK), Src, and Akt. Importantly, our results show the reduced expression of epithelial marker (E-cadherin) and induction of mesenchymal marker (N-cadherin) in HCV-infected cells. We also show the migration and invasion of HCV-infected cells using wound healing assay and matrigel coated Boyden chamber. In addition, we demonstrate the activation of above EMT markers, and the critical players involved in OPN-mediated cell signaling cascade using primary human hepatocytes infected with Japanese fulminant hepatitis (JFH)-1 HCV. Taken together, these studies suggest a potential role of OPN in inducing chronic liver disease and HCC associated with chronic HCV infection.
    PLoS ONE 01/2014; 9(1):e87464. DOI:10.1371/journal.pone.0087464 · 3.23 Impact Factor
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